PL178791B1 - Novel cyclopeptides and method of obtaining them as well as pharmaceutical preparation and method of obtaining same - Google Patents

Abstract

Claimed are cyclopeptides of the formula cyclo-(Arg-B-Asp-D-E) (I) where B = Gly, Ala, -HN-Q-CO- and D and E are independently Gly, -HN-Q-CO-, Ala, Asn, Asp, Asp(OR), Arg, 3-cyclohexyl alanine, Cys, Gln, Glu, His, Ile, Leu, Lys, Lys(Ac), Lys (AcNH2), Lys(AcSH), Met, 3-(2-naphthyl)-alanine, norleucine, ornithine, Phe, 4-halogen-Phe, phenylglycine, proline, 3-(2-pyridyl)-alanine, Ser, Thr, 3-(2-thienyl)-alanine, tetrahydroisoquinoline-3-carbonic acid, Trp, Tyr or Val or their derivs., R = alkyl with 1-6 C-atoms, Hal = F, Cl, Br, I; Q = alkylene with 1-6 C atoms and Ac = alkanoyl with 1-10 C-atoms, where applicable either the D- and L-forms, and the physiological salts thereof. Also claimed is (a) the prodn. of I, (b) a pharmaceutical compsn. including I, and (c) (b's) prodn.

Description

The present invention relates to novel compounds, cyclopeptides and their salts, and to the preparation of these new compounds. These new substances are cyclic adhesion inhibitors.

The new cyclopeptides in free form or in the form of pharmacologically acceptable salts are embraced by Formula 1, wherein B is Gly, Ala, -HN-Q-CO- and each of the symbols D and E is independently Gly, -HN-Q-CO -, Ala, Asn, Asp, Asp (OR), Arg, Cha, Cys, Gin, Glu, His, Ile, Leu, Lys, Lys (Ac), Lys (AcNH ₂ ), Lys (AcSH), Met, Nal , Nle, Om, Phe, 4-Hal-Phe, Phg, Pro, Pya, Ser, Thr, Tia, Tic, Trp, Tyr or Val, and the specified amino acid radicals can also be derivatized, R is 1- alkyl 6 carbon atoms, Hal is F, Cl, Br or J, Q is alkylene of 1-6 carbon atoms, and Ac is alkanoyl of 1-10 carbon atoms, the radicals of optically active amino acids and amino acid derivatives, both -D and -L variants are included.

Similar compounds are known from: Pharmazie 40, (8), 532-5 (1985).

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The invention is based on the task of developing new compounds with valuable properties, especially those that can be used in the preparation of medicaments.

It has been found that the compounds of formula I and their salts exhibit very valuable pharmacological properties. Above all, they act as integrin inhibitors, where they particularly inhibit the interaction of integrin receptors or Pj _-β-5 ligands. These compounds are particularly effective in the case of integrin <ι _ν β ₃ , ajf ia and _Ilb e ₃ . This effect can be detected, for example, according to a method that is described by JW Smith et al. In J. Biol. Chem. 265, 1267-12271 (1990). Additionally, there are anti-inflammatory effects. All these effects can be detected by methods known from the literature.

The compounds of the formula I can be used as active substances for medicaments in human and veterinary medicine, in particular for the prevention and treatment of circulatory diseases, thrombosis, myocardial infarction, arteriosclerosis, inflammation, stroke, angina, neoplastic diseases, osteolytic diseases, in particular osteoporosis, vascular development and restenosis after vascular plastic surgery. Moreover, the compounds can be used to improve the healing of wounds.

These compounds are suitable as antimicrobial active substances for the prevention of infections which could be caused by bacteria, fungi or yeasts. These substances can therefore advantageously be added as accompanying antimicrobial active substances when treating organisms into which substances that are foreign to the body are introduced, for example biological materials, implants, catheters or pacemakers. These substances act as antiseptics.

The previously and subsequently listed amino acid radical abbreviations are those of the following acids:

Abu 4-aminobutyric acid Oh 6-aminocaproic acid Ala alanine Asn asparagine Asp aspartic acid Asp (OR) β-ester of aspartic acid Arg arginine Cha 3-cyclohexylalanine Cit citrulline Cys cysteine Oak 2,4-diaminobutyric acid Main glutamine Glu glutamic acid Gly glycine His histidine How much isoleucine Leu leucine Lys lysine Lys (Ac) N ^E -alkanoyl lysine Lys (AcNH ₂ ) N + aminoalkanoyl lysine Lys (AcSH) N + mercaptoalkanoyl lysine Underworld methionine Nal 3 - (2-naphthyl) -alanine Nle norleucine Ohm omitin Phe phenylalanine 4-Hal-Phe 4-halo-phenylalanine Phg phenylglycine Pro proline

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Pya 3- (2-pyridyl) -alanine Cheese serine Yee 3- (2-thienyl) -alanine Tic tetrahydroisoquinoline-3 carboxylic acid Thr threonine Trp tryptophan Tyr tyrosine Yal valine.

The further abbreviations mean the following:

BOC III-row -butoxycarbonyl

CBZ benzyloxycarbonyl

DCCI is dicyclohexylcarbodiimide

DMF dimethylformamide

EDCI N-ethyl-N '- (3-dimethylaminopropyl) -carbodiimide hydrochloride

Et ethyl

Fmoc 9-fluorenylmethoxycarbonyl

HOBt 1-hydroxybenzotriazole

Me methyl

Mtr 4-methoxy-2,3,6-trimethylphenylsulfonyl

Obut tertiary butyl ester

Ome methyl ester

Oet ethyl ester

Phenoxyacetyl POA

TBTU 2- (1H-benzofriazolyl-1) -1,1,3,3-louronium tetrafluoroborate

TFA trifluoroacetic acid.

While the aforementioned amino acids can exist in several enantiomeric forms, previously and then, e.g. as a component of formula 1, all these forms as well as mixtures thereof (e.g. DL-variants) are included in addition to amino acids, e.g. as a component of the compounds of formula 1 1 may be equipped with suitable protecting groups known to each other.

The invention furthermore relates to a process for the preparation of new cyclopeptides of formula I, in which all symbols have the meanings given for formula 1, and their salts, according to the invention in that the compound of formula I is released from its functional function by treatment with a solvolizing or hydrogenolizing agent. derivative or that a peptide of formula 2, wherein Z is -Arg-B-Asp-DE-, -B-Asp-DE-Arg-, -Asp-DE-Arg-B-, -DE-Arg-B- Asp- or -E-Arg-B-Asp-D-, or a reactive derivative of such a peptide, is treated with a cyclizing agent, and / or that a basic or acidic compound of formula I is converted into one of its salts by treatment with an acid or a base .

Previously and thereafter, the symbols B, D, E, and Z have the meanings given when discussing Formulas 1 and 2, unless expressly stated otherwise.

In the preceding formulas, alkyl is preferably methyl, ethyl, isopropyl or tert-butyl.

The symbol B is preferably Gly, and also -HN-f-CIEG-CO-, -HN- (CH2) 3-CO-, -HN- (CH2) ₅ -CO- or Ala. The D symbol is preferably Phe, especially D-Phe, also 4-Hal-Pht, in particular 4-I-Phe and Pro, Tic, Lys, Nal or Phg, with the D-variants being particularly preferred.

Accordingly, the invention relates in particular to those compounds of formula I in which at least one of the radicals cited has one of the preferred meanings given above.

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A preferred set of compounds can be expressed by the arbitrary partial formula Ia given below, which properly corresponds to formula 1 but where B is Gly, -HN- (CH ₂ ) ₂ -CO-, -HN- (CH ₂ ) ₃ -CO -, -HN- (CH ₂ ) ₅ -CO- or Ala, D is D-Pro, D-Tic, Phe, D-Nal, D-Phg or 4-J-Phe and E is Val, Lys, Gly , Ala, Phe, Leu, Lys (Ac) or No.

A further preferred set of compounds can be expressed by the arbitrary partial formula 1b given below, which corresponds properly to formula 1 but where B is Gly, D is D-Phe and E is Val, Lys or Gly.

The compounds of formula I as well as the starting materials for their preparation are generally prepared analogously to known methods, such as those described in the literature (e.g. in standard publications such as e.g. Houben-Weyl, Methoden der Organischen Chemie, Georg-Thieme-Verlag, Stuttgart), namely, under reaction conditions which are known and suitable for the discussed reactions. It is also possible to use known variants of the procedure which are not mentioned in more detail here.

If desired, the starting materials can optionally also be prepared in situ in such a way that they are not separated from the reaction mixture, but are immediately further reacted to give compounds of formula I.

The compounds of formula I can be obtained by liberating them from their functional derivatives by solvolysis, in particular hydrolysis, or by hydrogenolysis.

Preferred substrates for solvolysis or hydrogenolysis are those compounds which, instead of one or more free amino and / or hydroxyl groups, contain appropriate protected amino and / or hydroxyl groups, preferably those compounds which, instead of the H atom bound to the N atom, have an amine protecting group. especially those compounds which correspond properly to formula 1, but instead of the group -NLE, have a group -NH-R ', where R' is an amine protecting group, e.g. BOC or CBZ.

Preference is also given to substrates which have a hydroxyl protecting group instead of the H-atom of the hydroxyl group, e.g. those compounds which correspond to formula I properly, but which instead of the hydroxyphenyl group contain an RO-phenyl group, where R is a hydroxyl protecting group.

Multiple - identical or different - protected amino and / or hydroxyl groups can also be present in the molecule of this substrate. If the protecting groups present are different from one another, they can in many cases be cleaved off selectively.

The expression "amine protecting group" is generally known and extends to groups which are suitable for protecting (for blocking) an amino group against chemical reactions, but also which are easily cleaved once the desired reaction has been carried out elsewhere in the molecule. Such typical groups are especially unsubstituted or substituted acyl, aryl, aralkoxymethyl or aralkyl groups. Since the amine protecting groups are removed after the desired reaction (or a series of reactions), their nature and size are not limiting; however, groups having 1-20, especially 1-8 carbon atoms are preferred. In connection with the present method, the term "acyl group" is to be understood in the broadest sense. They include acyl groups derived from aliphatic, araliphatic, aromatic or heterocyclic carboxylic or sulfonic acids, and especially alkoxycarbonyl, aryloxycarbonyl and, above all, aralkoxycarbonyl groups. Examples of such acyl groups are alkanoyl such as acetyl, propionyl, butyryl; aralkanoyl such as phenylacetyl; aroyl such as benzoyl or toluoyl; aryloxyalkanoyl such as POA; alkoxycarbonyl such as methoxycarbonyl, ethoxycarbonyl, 2,2,2-trichlorethoxycarbonyl, BOC, 2-iodoethoxycarbonyl; aralkyloxycarbonyl, such as CBZ (benzyloxycarbonyl), 4-methoxybenzyloxycarbonyl, Fmoc; arylsulfonyl such as Mtr. Preferred amine protecting groups are BOC and Mtr, furthermore CBZ, Fmoc, benzyl and acetyl.

The expression "hydroxyl protecting group" is also generally known and extends to groups which are suitable for protecting the hydroxyl group against chemical reactions but also which are easily cleaved once the desired reaction has been carried out elsewhere in the molecule. Typical groups of this kind are the abovementioned unsubstituted or substituted aryl, aralkyl or acyl groups, and furthermore also alkyl groups. Character and greatness

These hydroxyl protecting groups are not intended to be limiting as these groups are removed after the desired reaction or series of reactions; groups with 1-20, especially 1-10 carbon atoms are preferred. Examples of hydroxyl protecting groups are. in. benzyl, p-nitrobenzoyl, p-toluenesulfonyl, tert-butyl and acetyl, with benzyl and tert-butyl being particularly preferred. The COOH groups in aspartic and glutamic acid are preferably protected in the form of their tert-butyl esters (e.g. Asp (OBut)).

Derivatives of the compounds of formula I used as functional substrate may be prepared by known amino acid and peptide synthesis methods, such as described, for example, in the referenced standard works and patent applications, e.g. by the Merrifield solid phase method (BF Gysin and RB Merrifield, J. Am Chem. Soc. 94, page 3102 et seq. (1972)).

The liberation of the compounds of formula I from their functional derivatives is possible - depending on the protecting group used - e.g. with strong acids, preferably with TFA or perchloric acid, but also with other strong inorganic acids, such as hydrochloric or sulfuric acid, strong carboxylic acids, such as trichloroacetic acid, or sulfonic acids such as benzene or p-toluenesulfonic acid. The presence of an additional inert solvent is possible but not always needed. Suitable inert solvents are preferably organic solvents, for example carboxylic acids such as acetic acid, ethers such as tetrahydrofuran or dioxane, amides such as DMF, halogenated hydrocarbons such as dichloromethane and also alcohols such as methanol, ethanol or isopropanol. , and water. Mixtures of the abovementioned solvents are also possible. TFA is preferably used in excess without addition of further solvent, perchloric acid is preferably used in the form of a 9: 1 mixture of acetic acid and 70% perchloric acid. The temperature of the cleavage reaction is conveniently in the range of about 0-50 ° C; preferably the procedure is carried out at a temperature of 15-30 ° C (room temperature).

The BOC, OBut and Mtr groups can preferably be cleaved with TFA in dichloromethane or with about 3-5 n HCl in dioxane at 15-30 ° C, the FMOC group with about 5-20% dimethylamine, diethylamine or piperidine in DMF at 15-30 ° C.

Hydrogenolytically cleavable protecting groups (e.g. CBZ or benzyl) can be cleaved e.g. by treatment with hydrogen in the presence of a catalyst (e.g. a noble metal catalyst such as palladium, preferably on a support such as carbon). Suitable solvents in this connection are the solvents mentioned above, in particular, for example, alcohols, such as methanol or ethanol, or amides, such as DMF. As a rule, this hydrogenolysis is carried out at a temperature of about 0-100 ° C and a pressure of about 0.1-20 MPa, preferably at a temperature of 20-30 ° C and a pressure of 0.1-1 MPa. Hydrogenolysis of the CBZ group is readily accomplished, for example, on a 5-10% carbon-supported Pd catalyst in methanol medium or with ammonium formate (instead of H 2) on Pd-C catalyst in methanol / DMF system at 20-30 ° C.

Compounds of formula I can also be obtained by cyclization of compounds of formula II under the conditions of peptide synthesis. In this connection, it is expediently carried out by known peptide synthesis methods, such as described, for example, in Houben-Weyl, 1.c., vol. 15 / II, pages 1-86 (1974).

This reaction preferably takes place in the presence of a dehydrating agent, e.g. a carbodiimide such as DCCl or EDCl, moreover propanephosphonic acid anhydride (cf. Angew. Chem. 92, 129 (1980), diphenylphosphoryl azide or 1,2-ethoxy-N-ethoxycarbonyl-1,2 - dihydroquinoline, in an inert solvent, e.g. a halogenated hydrocarbon such as dichloromethane, an ether such as tetrahydrofuran or dioxane, an amide such as DMF or dimethylacetamide, a nitrile such as acetonitrile at a temperature from about -10 ° C to about 40 DEG C., preferably at a temperature of 0-30 DEG C. In order to accelerate the intramolecular cyclization before the intramolecular peptide bond, it is expedient to proceed in dilute solutions (idea of dilution).

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Instead of the compounds of the formula II, it is also possible to use corresponding reactive derivatives of these substances in this reaction, for example compounds in which reactive groups are transiently protected with protecting groups. The acid derivatives of formula II can be used, for example, in the form of their activated esters, which are deliberately formed in situ, for example by adding HOBt or N-hydroxysuccinimide.

As a rule, the starting materials of the formula II are known. They can be prepared by known methods, e.g. the above-discussed peptide synthesis and cleavage of protecting groups.

As a rule, protected pentapeptides of the formula R'-Z-OR ', e.g. BOC-Z-OMe or BOC-Z-OEt, are first synthesized and first saponified to acids of the formula R'-Z-OH e.g. BOC-Z -OH; of these, the protective group R 'is cleaved off, thereby obtaining free peptides of formula II.

A base of the formula I can be converted with an acid into the corresponding acid addition salt. Suitable acids for this reaction are, in particular, acids which result in physiologically acceptable salts. Thus, inorganic acids, e.g. sulfuric acid, nitric acid, hydrohalic acids, such as hydrochloric or hydrobromic acid, phosphoric acids, such as orthophosphoric acid, sulfamic acid, and also organic acids, especially aliphatic, alicyclic, araliphatic, aromatic or heterocyclic acids, can be used. - or polybasic carboxylic, sulphonic or sulfuric acids, e.g. formic, acetic, propionic, pivalic, diethylacetic, malonic, succinic, pimelic, fumaric, maleic, lactic, tartaric, malic, benzoic, salicylic, 2- or 3-phenylpropionic acids, citric, gluconic, ascorbic, nicotinic, isonicotinic, methanol or ethanesulfonic, ethane bisulfonic, 2-hydroxyethanesulfonic, benzenesulfonic, p-toluenesulfonic, naphthalene-mono- and-di-sulfonic, laurylsulfonic. Salts with physiologically non-neutral acids, e.g. picrates, can be used to isolate and / or purify compounds of formula I.

The acid of the formula I can be converted into one of its physiologically acceptable metal or ammonium salts by reaction with a base. Suitable salts are, in particular, the sodium, potassium, magnesium, calcium and ammonium salts, and moreover substituted ammonium salts, e.g. dimethyl, diethyl or diisopropyl ammonium salts, monoethanol, diethanol or triethanolammonium salts, cyclohexyl or dicyclohexylammonium, dibenzylethylenediammonium salts and further, for example, for example, salts with N-methyl-D-glucamine or with arginine or lysine.

The novel compounds of the formula I and their physiologically acceptable salts can be used for the production of pharmaceutical preparations by converting them together with at least one excipient or auxiliary substance and, if desired, together with one or more other active substances, into a suitable dosage form. . The preparations thus obtained can be used as medicaments in human and veterinary medicine. Suitable carrier substances are organic or inorganic substances which are suitable for enteral (e.g. oral or rectal), parenteral (e.g. intravenous injection) or topical (e.g. punctual, dermal, ophthalmic or nasal) application or inhalation aerosol application. and do not react with these new compounds, for example water or aqueous isotonic sodium chloride solution, lower alcohols, vegetable oils, benzyl alcohols, polyethylene glycols, glycerol triacetate and other fatty acid glycerides, gelatin, soy lecithin, carbohydrates such as lactase or starch , magnesium stearate, talc, cellulose, petroleum jelly. For oral administration, in particular tablets, dragées, capsules, syrups, juices or drops are used; lacquered tablets and capsules with gastro-resistant coatings or coated capsules are of special interest. For rectal administration, suppositories are used, for parenteral administration, solutions, preferably oily or aqueous solutions, moreover, suspensions, emulsions or implants are used. For topical application, for example, solutions which can be used in the form of eye drops, and for example suspensions, emulsions, creams, ointments or compresses, are suitable. For use as an inhalation aerosol, sprays which contain the active ingredient either dissolved or suspended in an expansion gas or an expansion gas mixture (e.g. CO ₂ or fluorocarbons) can be used. This is used on purpose

The active ingredient is in micronized form, whereby one or more physiologically acceptable solvents, for example ethanol, may additionally be added. Inhalation solutions can be administered with ordinary inhalers. The novel compounds can also be lyophilized and the obtained lyophilizates used, for example, for the preparation of injectables. The injections can be administered by the bolus or as a continuous infusion (e.g. intravenously, intramuscularly, subcutaneously or intrathecally). The preparations mentioned can be sterilized and / or can contain auxiliary substances, such as preservatives, stabilizers and / or wetting agents, emulsifiers, salts for influencing the osmotic pressure, buffer substances, dyes and / or fragrances. If desired, they can also contain one or more other active ingredients, for example one or more vitamins.

The substances according to the invention are generally used analogously to other commercially available peptides, in particular analogously to the compounds described in US-A-4,472,305, preferably in doses of about 0.05 to 500 mg, in particular 0.5 to 100 mg. dosage unit. The daily dosage is preferably in the range of about 0.01-2 mg / kg body weight. The specific dose for each particular patient, however, depends on a wide variety of factors, for example the efficacy of the specific compound employed, age, body weight, general health, sex, diet, timing and method of administration, excretion rate, combination of drugs. and the severity of the particular condition that is being treated. Parenteral administration is preferred.

Moreover, the novel compounds of formula I can be used as integrin ligands for the preparation of specific sorption chromatography columns for the preparation of pure integrins.

Lignad, i.e. a derivative of the peptide of formula I, is covalently coupled via the handle functional groups on the polymeric support.

Polymeric solid phases known in peptide chemistry with advantageous hydrophytic properties, e.g. cross-linked polysaccharides such as cellulose, Sepharose or Sephadex®, acrylamides, polyethylene glycol-based polymers, polymers known as Tentakelpolymere®, are suitable as polymeric carrier materials. .

Suitable handle functional groups which are linked to polymeric supports are preferably linear alkylene chains with 2 to 12 carbon atoms which are directly bonded to the polymer at one end and which have a functional group at the other end, such as, for example, hydroxyl, amino, , mercapto, maleimide, or -COOH, and are suitable to link to the C- or N-terminus of a given peptide.

It is hereby possible for the peptide to be linked directly or via a second handle functional group to the polymer holder. It is further possible that peptides which contain amino acid radicals with functional side chains bind through them to a handle functional group of the polymer.

Also you can set radicals of amino acids that sąskładnikiem peptide of formula 1, modified in their side chains so as to be available to bond through, for example. A group-SH, -OH, -NH ₂ or -COOH, with the handle of the polymer.

In this case, common amino acids are possible, such as phenylalanine derivatives, which contain a mercapto-, hydroxy-, amino- or carboxyalkyl chain in the 4-position of the phenyl ring, the functional groups being at the end of the chain.

Examples of amino acid radicals whose side chain can directly serve as a handle functional group are e.g. Lys, Om, Arg, Dab, Asp, Asn, Glu, Gln, Ser, Thr, Cys, Cit or Tyr.

Examples of N-ended holders are radicals such as -CO-C _n H _2n -NH _2, -CO-C _n H _{2n OH;} -CO- _CnH2tl -SH or -CO-CJH6 -COOH n = 2-12, the length of the alkylene chain is not a limitation and these chains can also optionally be replaced, for example, by suitable aryl or alkylaryl radicals.

The C-terminated handle functional groups may be, for example, -O-CnH2n-SH, -O-CnH2n-OH, -O-Cn ^ -N ^, -O-Cn ^ -COOH, -NH-CnH2n-SH,

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-NH-C _n H2 _n -OH, -NH-C _n H _2n -NH ₂ or -NH-C _n H _2n -COOH, wherein n and the symbol of the alkyl chain is valid what is stated in the previous section.

N- and C-terminated handle groups can also serve as a handle building block for an already functional side chain of an amino acid radical. Examples include amino acid radicals such as Lys (CO-C ₅ H ₁₀ -NH2), Asp (NH-C ₃ H ₆ -COOH) or Cys (C ₃ H ₆ -NH2), whereby this handle is always linked to with a side chain functional group.

The preparation of the materials for the specific sorption chromatography for purification of integrin takes place under conditions which are customary for the condensation of amino acids and are also known and already mentioned in the preparation of the compounds of formula 1.

Previously and thereafter, all temperatures are given in degrees Celsius. In the following examples, the term "simple workup" means: add water, if necessary, neutralize, extract with diethyl ether or dichloromethane, separate, dry the organic layer over sodium sulfate, filter, evaporate and purify by chromatography on silica gel and / or by crystallization. RZ means the retention time (in minutes) in HPLC in a column with an agent called Lichrosorb® RP select B (250-4.7μ), circulation factor: 0.3% TFA in water; graduated isopropanol 0-80 vol.% in 50 minutes at 1 ml / min. Flow and detection at 215 nm. M + is the molecular peak in the mass spectrum obtained by the "fast atom bombardment" (FAB) method.

Example 1. A solution of 0.2 g H-Arg-Gly-Asp-D-Pro-Val-ONa [e.g. obtained from Fmoc-Arg-Gly-Asp-D-Pro-Val-O-Wang, where the group -O-Wang is the radical of the 4-hydroxymethyl-phenyloxymethyl-polystyrene resin used in the modified Merrifield techniques, by cleavage of the Fmoc with piperidine / DMF and cleavage of the resin with TFA / CIRCU (1: 1)] in 15 ml DMF is diluted with 85 ml dichloromethane and treated with 50 mg NaHCO ₃ . After cooling in a dry ice / acetone mixture, 40 µL of diphenylphosphoryl azide are added. After standing at room temperature for 16 hours, the solution was concentrated. This concentrate was subjected to gel permeation chromatography (Sephadex G-10 isopropanol column) (water 8: 2) and then purified as usual by HPLC. There is obtained cyclo- (ArgGly-Asp-D-Pro-Val); RT = 13.4; M + 525.

Similarly, by cyclization of appropriate linear peptides, the following is obtained:

cyclo- (Arg-Gly-Asp-D-Tic-Val); RR (11.4; M + 587; cyclo- (Arg-Gly-Asp-D-Phe-Val); M + 575; ^{cyclo -} (Ar ^g-Gly- Asp ^- D ^- Lys ^-Val); Noun - 6.8; M + 556; c ^YKL o ^- (Arcj-Gly-Asp-D-Plie- ^ ^ys); RR - 10.9; 604; cyclo- (Arg-Gly-Asp-D-Phe-Gly); RR - 1 (3.1; M + 533 _; C ^ykl o ^- (Ar ^g ~ ^Gl y- ^Asp - ^D_ph e ^- Al ^a); Noun - 14.3; M + 547; ^{cyclo -} (Ar ^gG l .A3p- ^{y-D-Ph ph} e ^e); RZ, - 22.0; M ⁺ 623; cyclo- (Arg-Gly-Asp-D-Phe-Leu); RR - 21.2; M + 589 _; cyclo- (Arg-Gly-Asp-D-Nal-Val); RR - 24.7; M + 625; cyclo- (Arg-Gly-Asp-D-Phg-Val); IR - 16.5; l4 561;

cyclo- (Arg-Gly-Asp-Phe-Gly); RT - 13.2; M + 533;

Cyclo- (TArg-Gly-Asp-D-Phe-Val); RT = 14.8; M + 547; cyclo- (Arg-Gly-Asp-Phe-D-Phe); RT = 20.2; IM + 623; cyclo- (Arg-Gly-Asp-Phe-D-Leu); RT = 21.4; M + 549; cyclo- (Arg (Mtr) -Gly-Atop-D-Phe-Lys); RT = 23.4; M + 812; cyclo- (Arg-Gly-Asp-D-Phe-Lys (H3C-CO); RZ = 17.0; M + 242; cyclo- (Arg (Mtr) -p-Ala-Asp-Phe-D4Val); RT = 28.2; M + 801; cyclo- (Arg-Gly-Asp-D-Phe-Nle); RT = 22.0; M + 589; cyclo- (D-Arg-Gly-D-Asp-D-Phe -Val); RT = 17.5; M + 575; cyclo- (Arg-Gly-E-Asp-D-Phe-D-Gly); RT = 18.7, 575;

cyclo- (Arg-D-Ala-Asp-I> Phe-Val); RT = 11.9; (M 558; cyclo- (Arg-D-Ala-Asp-Phe-D-Val); RT = 11; 1; (M 558; cyclo- (Arg-Aha-Asp-D-Phe-Val); RZ = 20.8; 1M 6231 cyclo- (Arg-Abu-Asp-Phe-D-Val); RZ = 17.2; 1M 620; cyclo- (Arg-Aha-Asp-Phe-D-Val); RZ = i ;, 8; 1M 6231 cyclo- (Arg-Abu-Asp-D-Phe-Val); RT = 17.8; fM 623;

cyclo- (Arg-Gly-Asp-D- (4-I-Phe) -Val); RT = 23.3; M + 701; cyclo- (Arg-Gly-Asp-Phe-Val); RT = 21.8; M + 575; cyclo- (Arg-Gly-D-Asp-D-Phe-Val); RT == 20.7; M + 575; cyclo- (D-Arg-Gly-Asp-Phe-D-Val); RT = 20.8; M ⁺ 555; cyclo- (D-Arg-Gly-Asp-D-Phe-Val); RT = 2 ^ 9, ^ M + 555; cyclo- (Arg-Gly-I> 7Sp-D-Phe-Val). R Z = 20.7; iR 555; cyclo- (Arg-Gly-Atop-Phe-E-Nal);

cyclo- (Arg-Gly-Asp-Phe-D-Leu);

cyclo- (Arg-Gly-Asp-Phe-D-Ser);

cyclo- (Arg-Gly-Asp-D-Nal-Leu);

cyclo- (Arg-Gly-Asp-Nal-D-Val (;

cyclo- (Arg-Gly-Asp-Phg-I-al);

cyclo- (Arg-Gly-Asp-Trp-D-Val);

Cyclo - ((Arg-Giy-Asp-Ala (2-thienyl) -D-Val);

cyclo- (Arg-Gly-Asp-Lys-D-Val);

cyclo- (Arg-Gly-Asp- (4-NC> 2-Phe) -D-Val);

cyclo- ((Arg-Gly-Asp-Cha-D-Val);

cyclo- (Arg-Gly-Asp-D-Phe-β-Ala);

cyclo- (Arg-Gly-Asp-D-Phe-Abu);

cyclo- ((Arg-Gly-Asp-D-Phe-Ahia);

cyclo- (Arg-Gly-Asp-D-Phe-Lys);

cyclo- (Arg-Gly-Asp-D-Phe-Pro);

cyclo- (Arg-Gly-Asp-D-Phe-Arg);

cyclo- (Arg-Gly-Asp-D-Phe-Lys (Aha));

cyclo- (Arg-Gly-Asp-D-Phe-MIal);

cyclo- (Arg-Gly-Asp-D-Phe-Ser);

cyclo- (D-Arg ((Mtr) -Gly-Asp (OBiut) -D-Phe-Val);

cyclo- (D-Arg (Mtr) -Gly-Asp (CBut) -D-Phe-Val);

cyclo- (Arg (Mtr) -Gly-D-Asp (OEt) -D-Phe-Val); cyclo- (Arg (Mtr) -Gly-Asp-Phe-D-Nal); cyclo- ((Arg (Mtr) -Gly-Asp-Phe-D-Leu); cyclo- ((Arg (Mtr) -Gi.y-Asp-Phe-D-Ser); cyclo- ((Arg ((Mtr ) -Gly-Asp-D-Nal-Leu); cyclo- ((Arg ((Mtr) -Gly-Asp-Nal-D-Val); cyclo- ((Arg ((Mtr) -Gly-Asp-Phg- D-Val); cyclo- (Arg (Mtr) -Gly-Asp-Trp-D-Val); cyclo- (Arg-Gly-Asp-D-Phe-Lys (BOC-Aha); cyclo- ((Arg- Gly-Asp-D-Phe-Lys (CO-CH2SH)); cyclo- (Arg-Gly-Asp-D-Phe-Lys (CC- (CH2) 2 SH)); cyclo- (Arg-Gly-Asp- D-Phe-Lys ((CO- (CH2) 3SH)); cyclo- (Arg-Gly-Asp-D-Phe-Lys (CC- (CH2) 4SH)); cyclo- ((Arg-Gly-Asp- D-Phe-Lys ((CC- (CH2) 5SH));

Cyclo- (Arg-Gly-Asp-I> iyr-Val); cyclo- (Arg-Gly-Asp-D- (4-Cl-Phe) -Val); cyclo- (Arg-Gly-Asp-D-Eya-Val); cyclo- (Arg-Gly-Asp-D-Tyr (OEt) -Val); cyclo- (Arg-Gly-Asp-D-Arg-Val); cyclo- (Arg-Gly-Asp-D-Trp-Val); cyclo- (Arg-Gly-Asp-D-Ala (2-thienyl) -Val); cyclo- (Arg-Gly-Asp-D-Tyr-NMe-Val); cyclo- (Arg-Gly - D-Asp-D-Phe-Val); cyclo- (Arg-Gly-D-Asp-Phe-D-Val); cyclo- (D-Arg-Gly-Asp-Phe-D-Val); cyclo- (Arg-Gly-D-Asp-Phe-P - Val).

Example II. A solution of 0.28 g of cyclo- (Arg- (Mtr) - [3-Ala-Asp-Phe-D-Val) [obtained by cyclization according to example 1] in 8.4 ml of TFA, 1.7 ml of dichloromethane and 0 9 ml of thiophenol are left for 4 hours at room temperature, then concentrated and, after dilution with water, freeze-dried. Gel chromatography on a medium named Sephadex G 10 (acetic acid / water 1: 1) and subsequent purification by preparative HPLC under the indicated conditions gives cyclo- [Arg-p-Ala-Asp-Phe-D-Val) ; Rz = 17.0; M + 589.

Similarly:

from cyclo- (Arg (Mtr) -Gly-Asp-D-Phe-Iys) gives cyclo- (Arg-Gly-Asp-D-Phe-Lys); RT = 10.9; M ⁴ 604;

from cyclo- (I ^^ - Arg (Mtr) -Gly-Asp (OBut) -D-Phe-Val) gives cyclo- (D-Arg-Gly-Asp-D-Phe-Val);

from cyclo- (Arg (Mtr) -Gly-D-7Asp (OEt) -D-Phe-Val) gives cyclo- (Arg-Gly-D-Asp-D-Phe-Val);

from cyclo- (Arg (Mtr) -Gly-Asp-Phe-1-Nal) gives cyclo- (Ar-g-Gly-Asp-Phe-D-Nal);

from cyclo- (A.rgIMtr) -Gly-Asp-Phe-I-Leu) gives cyclo- (Arg-Gly-Asp-Phe-D-Ieu);

From cyclo- (Arg (Mtr) -Gly-Asp-Phe-I> -Ser) gives cyclo- (Arg-Gly-Asp-Phe-D-Ser);

from cyclo- (Arg (Mtr) -Gly-Asp-D-Nal-Leu) -Gly-Asp-D-Nal-Leu); z cyclo- ((Arg ((Mtr) -Gly-Asp-Nal-D-Val) -Gly-Asp-Nal-D-Val); z cyclo- (Arg (Mtr) -Gly-Asp-Phg-D- Val) -Gly-Asp-Phg-D-Val); from cyclo- (Arg (Mtr) -Gly-Asp-Trp-D-Val) -Gly-Asp-Trp-I> -Val) is obtained cyclo- (Arresting cyclo- (Arresting cyclo- (Arresting cyclo- (ArgExample III. 80 mg of cyclo- (Arg-Gly-Asp-D-Phe-Leu) are dissolved five to six times in 0.01 m HCl and lyophilized after each dissolution process. Subsequent purification by HPLC gives become cyclo- (Arg-Gly-Asp-D-Phe-Leu) x HCl; RT = 20.6, M + 589.

Similarly:

from cyclo- (Arg-Gly-Asp-D-Phe-Val) gives cyclo- (Arg-Gly-Asp-D-Phe-Val) x HCl; RT = 18.4; M + 575;

from cyclo- (Arg-Gly-Asp-D-Phe-Leu) gives cyclo- (Arg-Gly-Asp-D-Phe-Leu) x HCl; cyclo- (Arg-Gly-Asp-D-Phe-Leu) by treatment with acetic acid affords cyclo- (Arg-Gly-Asp-D-Phe-Leu) x H3C-COOH; RT = 19.2: M + 589;

cyclo- (Arg-Gly-Asp-D-Phe-Leu) by treatment with nitric acid affords cyclo- (Arg-Gly-Asp-D-Phe-Leu) x HNO ₃ ; RT = 20.4; M + 589.

Example IV. In order to prepare specific sorption phases, 0.9 g of N-maleimide- (CH ₂ ) ₅ -CO-NH- (CH2) ₃ -polymer [obtained by condensation of N-meleinimide- (CH2) 5-COOH with H2N- (CH2) _3- polymer] in 10 ml 0.1M sodium phosphate buffer at pH = 7 and the total is added at 4 ° C to the equivalent of cyclo- (Arg-Gly-Asp-D-Phe-Lys (CO (CH2 ) 2SH)). The mixture is stirred for 4 hours while heating the reaction mixture to room temperature, the solid residue is filtered off and washed twice with 10 ml of buffer solution (pH = 7) and then washed three times with 10 ml of water each time. Cyclo- (Arg-Gly-Asp-D-Phe-Lys (CO (CH2) 2. -S-3- (N-maleimide- (CH2) 5-CONH- (CH2) 3-polymer)) is obtained.

Example 5 Analogously to example I, by condensation of the substance polymer-O- (CH ₂ ) 3-NH2 [commercially available] and cyclo (Arg-Gly-Asp-D-Pro-Lys (CO (CH ₂ ) 4COOH [obtained by condensation of adipic acid with cyclo- (Arg-Gly-Asp-D-Pro-Lys) under the given conditions] the following polymeric phase is obtained: cyclo- (Arg-Gly-Asp-DPro-Lys- (CO- (CH2) 4-CO-NH- (CH ₂ ) 3-O-polymer).

Analogously, the condensation of cyclo- (Arg-Gly-Asp-D-Phe-Lys- (CO- (CH2) 5-NH ₂ )) with HOOC-CH2-O-polymer gives cyclo- (Arg-Gly-Asp-D -Phe-Lys- (CO- (CH2) 5-NH-CO-CH ₂ -O-polymer)).

The following examples relate to pharmaceutical preparations.

Example VI. Injection vials

A solution of 100 g of cyclopeptide I and 5 g of disodium hydrogen phosphate in 3 liters of doubly distilled water is adjusted with 2N hydrochloric acid to a pH of

178 791 pH = 6.5, filter under sterile conditions, fill vials with it, lyophilize under sterile conditions and sterile seal. Each vial contains 5 mg of the active ingredient.

Example VII. Suppositories

A mixture of 20 g of the active ingredient of the formula I is melted with 100 g of soy lecithin and 1400 g of cocoa butter, poured into molds and allowed to solidify. Each suppository contains 10 mg of active ingredient.

Example VIII. Solution

A solution of 1 g of the active ingredient of the formula 1.9.38 g of Na [hPO .., x 2H2O, 28.48 g of Na2HPO4 x 12H2O and 0.1 g of benzalkonium chloride in 940 ml of doubly distilled water is prepared. The pH of the solution is adjusted to 6.8, made up to 1 liter and sterilized by irradiation. This solution can be used as eye drops.

Example IX. Ointment

500 mg of active ingredient of the formula I are mixed with 99.5 g of petroleum jelly under aseptic conditions.

Example X. Tablets

A mixture of 100 g of cyclopeptide of formula 1.1 kg of Lokatose, 600 g of microcrystalline cellulose, 600 g of maize starch, 100 g of polyvinylpyrrolidone, 80 g of talc and 10 g of magnesium stearate is compressed into a tablet as usual, so that each tablet contains 10 mg of the substance active.

Example XI. Dragees

In analogy with Example 10, tablets are pressed and then coated in a known manner with sucrose, corn starch, talc, tragacanth and dye.

Example XII. Capsules

Hard gelatin capsules are filled in the usual manner with the active ingredient of the formula I such that each capsule contains 5 mg of active ingredient.

Example XIII. An inhalation aerosol of g of the active compound of the formula I is dissolved in 10 liters of isotonic sodium chloride solution and this solution is filled into commercially available aerosol canisters with an injection mechanism. This solution can be sprayed into the mouth or nose. One spray (approximately 0.1 ml) corresponds to a dose of approximately 0.14 mg.

Cyclo- (Arg-B-Asp-D-E

Formula 1

H-Z-OH

Formula 2

Publishing Department of the UP RP. Circulation of 70 copies

Price PLN 4.00.

Claims (4)

Patent claims 1. New compounds, cyclopeptides of formula I, in which B represents Gly, Ala, -HN-Q-CO- and each of the symbols D and E independently of each other represents Gly, -HN-Q-CO-, Ala, Asn, Asp, Asp (OR), Arg, Cha, Cys, Gln, Glu, His, Ile, Leu, Lys, Lys (Ac), Lys (AcNH ₂ ), Lys (AcSH), Met, Nal, Nle, Om, Phe , 4-Hal-Phe, Phg, Pro, Pya, Ser, Thr, Tia, Tic, Trp, Tyr or Val, and the specified amino acid radicals can also be derivatized, R is an alkyl of 1-6 carbon atoms, Hal is F, Cl, Br or J, Q is alkylene of 1-6 carbon atoms and Ac is alkanoyl of 1-10 carbon atoms, where both the radicals of optically active amino acids and amino acid derivatives are included both D-forms and -L, and the pharmacologically acceptable salts of these cyclopeptides. 2. A compound according to claim 1. which is (a) cyclo- (Arg-Gly-Asp-D-Lys-Val); (b) cyclo- (Arg-Gly-Asp-D-Phe-Lys); (c) cyclo- (Arg-Gly-Asp-D-Phe-Gly); (d) cyclo- (Arg-Gly-Asp-D-Phe-Phe); (e) cyclo- (Arg-Gly-Asp-D-Phe-Leu); (f) cyclo- (Arg-Gly-Asp-Phe-D-Leu). 3. A method for the preparation of new cyclopeptides of formula 1, wherein B is Gly, Ala, -HN-Q-CO- and each of the symbols D and E independently of each other represents Gly, -HN-Q-CO-, Ala, Asn, Asp, Asp (OR), Arg, Cha, Cys, Gln, Glu, His, Ile, Leu, Lys, Lys (Ac), Lys (AcNH2), Lys (AcSH), Met, Nal, Nle, Om, Phe, 4-Hal-Phe, Phg, Pro, Pya, Ser, Thr, Tia, Tic, Trp, Tyr or Val, and the specified amino acid radicals can also be derivatized, R is an alkyl of 1-6 carbon atoms, Hal is F , Cl, Br or J, Q is alkylene of 1-6 carbon atoms and Ac is alkanoyl of 1-10 carbon atoms, where both the radicals of optically active amino acids and amino acid derivatives are both D and and -L, and salts of these cyclopeptides, characterized in that by treatment with a solvolizing or hydrogenolizing agent the compound of formula 1 is released from a functional derivative thereof, or that a peptide of formula 2 in which Z is -Arg-B-Asp-DE- , -B-Asp-DE-Arg-, -Asp-DE-Arg-B-, -DE-Arg-B-Asp- or -E-Arg-B-Asp-D-, or a reactive derivative of such a peptide, is treated with the agent cyclizing, and / or that a basic or acidic compound of formula I is converted into one of its salts by treatment with an acid or base.